Table 1.

Oligonucleotides used for overlapping PCRs

Mutant	Mutation(s)a	Oligonucleotide
		Orientationb	Sequence
M1	R279 K282	For	GGGAAAGCCATAGAAGCAATAAAGCAAGAGC
		Rev	GCTCTTGCTTTATTGCTTCTATTGCTTTCCCAATTATATC
M2	K300	For	ACCAAGACCACCCATACGCCACGTGGGCATAC
		Rev	GTATGCCCACGTGGCGTATGGGTGGTCTTGGT
M3	K311	For	ATGGTAGCTATGAAACAGCACAGACTGGATCAGCA
		Rev	TGCTGATCCAGTCTGTGCTGTTTCATAGCTACCAT
M4	R325	For	CAACGGAGTGGTCGCGCTGCTGACAAAACCTTG
		Rev	CAAGGTTTTGTCAGCAGCGCGACCACTCCGTTG
M5	R361 K370	For	AAAGTGGACACGGCAACCCAAGAACCGAAAGAAGGCACGGCCAAACTAATGAAAA
		Rev	TTTTCATTAGTTTGGCCGTGCCTTCTTTCGGTTCTTGGGTTGCCGTGTCCACTTT
M6	K386 K388	For	AAAGAATTAGGGGCGAAAGCGACACCCAGGATGTGCACCAGAGAA
		Rev	TTCTCTGGTGCACATCCTGGGTGTCGCTTTCGCCCCTAATTCTTT
M7	R391 R395	For	GAAGAAAAAGACACCCGCGATGTGCACCGCAGAAGAATTCACAAG
		Rev	CTTGTGAATTCTTCTGCGGTGCACATCGCGGGTGTCTTTTTCTTC
M8	R400 K401	For	CCAGAGAAGAATTCACAGCAGCCGTGAGAAGCAATGC
		Rev	GCATTGCTTCTCACGGCTGCTGTGAATTCTTCTCTGG
M9	R438	For	TGGTTGACAAGGAAGCGAATCTCCATCTTGAAGG
		Rev	CCTTCAAGATGGAGATTCGCTTCCTTGTCAACCA
M10	K456 R457	For	ACATGATGGGAGCAGCTGAGAAGAAGCTAGGGGAATTCGGCAAGGC
		Rev	GCCTTGCCGAATTCCCCTAGCTTCTTCTCAGCTGCTCCCATCATGT
M10.1	K456	For	ACATGATGGGAGCAAGAGAGAAGAAGCTAGGGGAATTCGGCAAGGC
		Rev	GCCTTGCCGAATTCCCCTAGCTTCTTCTCTCTTGCTCCCATCATGT
M10.2	R457	For	ACATGATGGGAAAAGCTGAGAAGAAGCTAGGGGAATTCGGCAAGGC
		Rev	GCCTTGCCGAATTCCCCTAGCTTCTTCTCAGCTTTTCCCATCATGT
M11	K456 R457 K459 K460	For	TACAACATGATGGGAGCAGCTGAGGCAGCACTAGGGGAATTCGGCA
		Rev	TGCCGAATTCCCCTAGTGCTGCCTCAGCTGCTCCCATCATGTTGTA
M12	R481	For	TGTGGCTTGGAGCAGCCTTCTTAGAGTTTGAAGCCCTAGGAT
		Rev	ATCCTAGGGCTTCAAACTCTAAGAAGGCTGCTCCAAGCCACA
M13	R519 K523	For	GTTACATTCTAGCAGACGTGAGCGCAAAAGAGGGAGGAGCA
		Rev	TGCTCCTCCCTCTTTTGCGCTCACGTCTGCTAGAATGTAAC
M14	R540	For	CCGCAGGATGGGATACAGCAATCACACTAGAAGACCTAAAAAATG
		Rev	CATTTTTTAGGTCTTCTAGTGTGATTGCTGTATCCCATCCTG
M15	K561 K562	For	ATGGAAGGAGAACACGCAGCGCTAGCCGAGGCCATTTTCAAACT
		Rev	AGTTTGAAAATGGCCTCGGCTAGCGCTGCGTGTTCTCCTTCCAT
M16	K575 R578 R581	For	GTACCAAAACGCGGTGGTGGCTGTGCAAGCACCAACACCAAGA
		Rev	TCTTGGTGTTGGTGCTTGCACAGCCACCACCGCGTTTTGGTAC
M17	R648 R651	For	GCAAAACTGGTTAGCAGCTGTGGGGGCCGAAAGGTTATC
		Rev	GATAACCTTTCGGCCCCCACAGCTGCTAACCAGTTTTGC
M18	R769	For	ACTTCCACAGAGCCGACCTCAGGCTGGCGGCAAATGCTAT
		Rev	ATAGCATTTGCCGCCAGCCTGAGGTCGGCTCTGTGGAAGT
M19	K840 R841	For	AGGAAATCCCATACTTGGGGGCAGCGGAAGACCAATGG
		Rev	CCATTGGTCTTCCGCTGCCCCCAAGTATGGGATTTCCT

^aAll residues were mutated to alanines.

^bFor, forward; Rev, reverse.